Development and validation of a selective and effective pressurized liquid extraction followed by liquid chromatography-mass spectrometry method for the determination of fructosazine analogues in the ammonia treated extract of Eugenia jambolana Lamarck seeds.

This study describes a selective and effective pressurized liquid extraction (PLE) coupled with HPLC-DAD-ESI/MS method for the identification and quantification of three fructosazine analogues (FZAs), fructosazine, 2,6- and 2,5-deoxyfructosazine in Madeglucyl® (MG) which is an ammonia treated extract of Eugenia jambolana Lamarck seeds, and is the world's first anti-diabetic phytodrug. FZAs were extracted from MG by PLE using methanol as extraction solvent. The PLE extract was then analyzed directly by HPLC-DAD-ESI/MS without cleanup step. Chromatographic separation of these highly related structures was achieved on a porous graphic carbon (PGC) column. The identification of the target FZAs was confirmed by the similar retention time, similar UV and MS spectra to the corresponding pure standards. The quantification was performed by using an electrospray positive ionization mass spectrometry in the selected ion monitoring (SIM) mode. The PLE procedure was optimized and overall method was validated in terms of sensitivity, linearity, selectivity and matrix effect, precision, accuracy and recovery, and stability of the target FZAs in the aqueous solution and in the PLE extracts solution of MG. The developed method was proved to be selective, sensitive, precise, accurate for the quantification of FZAs in MG.

Pressurized liquid extraction and HPLC quantification of folic acid in fortified wheat flours.

A pressurized liquid extraction (PLE) method using phosphate buffer as solvent was applied for folic acid (FA) extraction from fortified wheat flours and was compared to a standard solid-liquid extraction (SLE) method. Extracted FA was quantified by reverse phase high-performance liquid chromatography (RP-HPLC) hyphenated with a phenyl column and an absorption photometric detector (λ = 280 nm). Detection and quantification limits were 0.12 and 0.4 ng, respectively, corresponding to 0.06 and 0.2 µg g(-1) of analyzed wheat flour. Equivalent FA contents were found by both extraction methods, but a single PLE allowed a total recovery of FA content, whereas at least three successive SLEs were needed to achieve a total recovery of FA. The obtained results indicated that PLE is a rapid and efficient technique for FA extraction from fortified wheat flour.